• 中國醫(yī)科大學附屬盛京醫(yī)院膽道、疝、減肥外科 (遼寧沈陽 110004);

目的  探討Ghrelin對大鼠離體胰腺組織胰島素分泌及葡萄糖轉運蛋白-2 (Glut-2)表達的影響。
方法  將25只Wistar大鼠隨機分為正常對照組(NC組)、高糖組(HCG組)、高糖+高濃度Ghrelin (10-8mol/L)組(HCG+HCGh組)、高糖+中濃度Ghrelin (10-9mol/L)組(HCG+MCGh組)及高糖+低濃度Ghrelin (10-10mol/L)組(HCG+LCGh組),每組5只。建立大鼠離體胰腺灌注模型,經(jīng)腹主動脈遠端相應灌注低糖 (5.5mmol/L)、單純高糖 (33.3mmol/L) 溶液或加入上述不同濃度Ghrelin的高糖 (33.3mmol/L) 溶液。采用ELISA法測定門靜脈流出液的胰島素水平,采用免疫組化染色方法半定量測定Glut-2蛋白在離體胰腺組織中的表達水平,采用透射電鏡觀察胰島β細胞的超微結構改變。
結果  5組大鼠的空腹血漿葡萄糖 (FBG)、空腹血漿胰島素(FINS)、胰島素抵抗指數(shù) (HOMA-IR) 及胰島素分泌指數(shù) (HOMA-β) 比較差異均無統(tǒng)計學意義 (P>0.05)。用低糖溶液灌注時,5組大鼠門靜脈流出液的胰島素水平比較差異無統(tǒng)計學意義 (P>0.05);而用高糖溶液灌注后,胰島素的分泌在3min和10~12min時出現(xiàn)了2個高峰 (以HCG+HCGh組最高)。NC組大鼠胰島β細胞Glut-2蛋白的平均光密度值高于其余4組 (P<0.05)。透射電鏡觀察結果顯示,高糖各組大鼠離體胰腺的凋亡征象較NC組嚴重,但HCG+HCGh組的細胞器損害程度較HCG+MCGh組及HCG+LCGh組輕。
結論  在大鼠離體胰腺中,Ghrelin可促進高濃度葡萄糖誘導的胰島素分泌,對胰島β細胞起保護作用。

引用本文: 于一凡,周勇,王勇,劉金鋼. Ghrelin對大鼠離體胰腺胰島素分泌及Glut-2蛋白表達的影響△. 中國普外基礎與臨床雜志, 2013, 20(3): 251-258. doi: 復制

1. Couzin J. Medicine. Bypassing medicine to treat diabetes[J]. Science, 2008, 320(5875):438-400.
2. Henquin JC, Ravier MA, Nenquin M, et al. Hierarchy of the beta-cell signals controlling insulin secretion[J]. Eur J ClinInvest, 2003, 33(9):742-750.
3. Wang Y, Liu J. Combination of bypassing stomach and vagus dissection in high-fat diet-induced obese rats—a long-term investigation[J]. Obes Surg, 2010, 20(3):375-379.
4. Irako T, Akamizu T, Hosoda H, et al. Ghrelin prevents development of diabetes at adult age in streptozotocin-treated newborn rats[J]. Diabetologia, 2006, 49(6):1264-1273.
5. Kojima M, Hosoda H, Date Y, et al. Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J]. Nature, 1999, 402(6762):656-660.
6. Mclaughlin T, Abbasi F, Lamendola C, et al. Plasma ghrelin concentrations are decreased in insulin-resistant obese adults relativeto equally obese insulin-sensitive controls[J]. Clin Endocrinol Metab, 2004, 89(4):1630-1635.
7. Pöykkö SM, Kellokoski E, Hörkkö S, et al. Low plasma ghrelin is associated with insulin resistance, hypertension, and the prevalence of type 2 diabetes[J]. Diabetes, 2003, 52(10):2546-2553.
8. 趙乃倩, 余葉蓉, 譚惠文, 等. 大鼠胰腺離體灌注技術的建立 [J]. 中華內分泌代謝雜志, 2007, 23(5):459-463.
9. Broglio F, Arvat E, Benso A, et al. Ghrelin,a natural GH secretagogue produced by the stomach,induces hyperglycemia andreduces insulin secretion in humans[J]. J Clin Endocrinol Metab,.
10. Egido EM, Rodriguez-Gallardo J, Silvestre RA, et al. Inhibitory effect of ghrelin on insulin and pancreatic somatostatin secretion[J]. Eur J Endocrinol, 2002, 146(2):241-244.
11. Reimer MK, Pacini G, Ahrén B. Dose-dependent inhibition by ghrelin of insulin secretion in the mouse[J]. Endocrinology, 2003, 144(3):916-921.
12. Date Y, Nakazato M, Hashiguchi S, et al. Ghrelin is present in pancreatic alpha-cells of humans and rats and stimulates insulin secretion[J]. Diabetes, 2002, 51(1):124-129.
13. Lee HM, Wang G, Englander EW, et al. Ghrelin, a new gastrointestinal endocrine peptide that stimulates insulin secretion:entericdistribution, ontogeny, influence of endocrine, and dietarymanipulations[J]. Endocrinology, 2002, 143(1):185-190.
14. Salehi A, Dornonville de la Cour C, Håkanson R, et al. Effects of ghrelin on insulin and glucagon secretion:a study of isolated pancreatic islets and intact mice[J]. Regul Pept, 2004, 118(3):143-150.
15. Chung H, Kim E, Lee DH, et al. Ghrelin inhibits apoptosis in hypothalamic neuronal cells during oxygen-glucose deprivation[J]. Endocrinology, 2007, 148(1):148-159.
16. Miao Y, Xia Q, Hou Z, et al. Ghrelin protects cortical neuron against focal ischemia/reperfusion in rats[J]. Biochem Biophys Res Commun, 2007, 359(3):795-800.
17. Thorens B, Wu YJ, Leahy JL, et al. The loss of GLUT2 expression by glucose-unresponsive beta cells of db/db mice is reversible and is induced by the diabetic environment[J]. J Clin Invest, 1992, 90(1):77-85.
18. Bell GI, Kayano T, Buse JB, et al. Molecular biology of mammalian glucose transporters[J]. Diabetes Care, 1990, 13(3):198-208.
19. Fukumoto H, Kayano T, Buse JB, et al. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues[J]. J Biol Chem, 1989, 264(14):7776-7779.
20. Thorens B, Cheng ZQ, Brown D, et al. Liver glucose transporter:a basolateral protein in hepatocytes and intestine and kidney cells[J]. Am J Physiol Cell Physiol, 1990, 259(6 Pt 1):C279-C285.
21. Thorens B, Sarkar HK, Kaback HR, et al. Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney, and beta-pancreatic islet cells[J]. Cell, 1988, 55(2):281-290.
22. Gould GW, Thomas HM, Jess TJ, et al. Expression of human glucose transporters in Xenopus oocytes:kinetic characterization and substrate specificities of the erythrocyte, liver, and brain isoforms[J]. Biochemistry, 1991, 30(21):5139-5145.
23. Orci L, Thorens B, Ravazzola M, et al. Localization of the pancreatic beta cell glucose transporter to specific plasma membrane domains[J]. Science, 1989, 245(4915):295-297.
24. Guillam MT, Hümmler E, Schaerer E, et al. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2[J]. Nat Genet, 1997, 17(3):327-330.
25. Guillam MT, Dupraz P, Thorens B. Glucose uptake,utilization,and signaling in GLUT2-null islets[J]. Diabetes, 2000, 49(9):1485-1491.
26. Guillemain G, Muñoz-Alonso MJ, Cassany A, et al. Karyopherin alpha2:a control step of glucose-sensitive gene expression in hepatic cells[J]. Biochem J, 2002, 364(Pt 1):201-209.
27. , 86(10):5083-5086.
  1. 1. Couzin J. Medicine. Bypassing medicine to treat diabetes[J]. Science, 2008, 320(5875):438-400.
  2. 2. Henquin JC, Ravier MA, Nenquin M, et al. Hierarchy of the beta-cell signals controlling insulin secretion[J]. Eur J ClinInvest, 2003, 33(9):742-750.
  3. 3. Wang Y, Liu J. Combination of bypassing stomach and vagus dissection in high-fat diet-induced obese rats—a long-term investigation[J]. Obes Surg, 2010, 20(3):375-379.
  4. 4. Irako T, Akamizu T, Hosoda H, et al. Ghrelin prevents development of diabetes at adult age in streptozotocin-treated newborn rats[J]. Diabetologia, 2006, 49(6):1264-1273.
  5. 5. Kojima M, Hosoda H, Date Y, et al. Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J]. Nature, 1999, 402(6762):656-660.
  6. 6. Mclaughlin T, Abbasi F, Lamendola C, et al. Plasma ghrelin concentrations are decreased in insulin-resistant obese adults relativeto equally obese insulin-sensitive controls[J]. Clin Endocrinol Metab, 2004, 89(4):1630-1635.
  7. 7. Pöykkö SM, Kellokoski E, Hörkkö S, et al. Low plasma ghrelin is associated with insulin resistance, hypertension, and the prevalence of type 2 diabetes[J]. Diabetes, 2003, 52(10):2546-2553.
  8. 8. 趙乃倩, 余葉蓉, 譚惠文, 等. 大鼠胰腺離體灌注技術的建立 [J]. 中華內分泌代謝雜志, 2007, 23(5):459-463.
  9. 9. Broglio F, Arvat E, Benso A, et al. Ghrelin,a natural GH secretagogue produced by the stomach,induces hyperglycemia andreduces insulin secretion in humans[J]. J Clin Endocrinol Metab,.
  10. 10. Egido EM, Rodriguez-Gallardo J, Silvestre RA, et al. Inhibitory effect of ghrelin on insulin and pancreatic somatostatin secretion[J]. Eur J Endocrinol, 2002, 146(2):241-244.
  11. 11. Reimer MK, Pacini G, Ahrén B. Dose-dependent inhibition by ghrelin of insulin secretion in the mouse[J]. Endocrinology, 2003, 144(3):916-921.
  12. 12. Date Y, Nakazato M, Hashiguchi S, et al. Ghrelin is present in pancreatic alpha-cells of humans and rats and stimulates insulin secretion[J]. Diabetes, 2002, 51(1):124-129.
  13. 13. Lee HM, Wang G, Englander EW, et al. Ghrelin, a new gastrointestinal endocrine peptide that stimulates insulin secretion:entericdistribution, ontogeny, influence of endocrine, and dietarymanipulations[J]. Endocrinology, 2002, 143(1):185-190.
  14. 14. Salehi A, Dornonville de la Cour C, Håkanson R, et al. Effects of ghrelin on insulin and glucagon secretion:a study of isolated pancreatic islets and intact mice[J]. Regul Pept, 2004, 118(3):143-150.
  15. 15. Chung H, Kim E, Lee DH, et al. Ghrelin inhibits apoptosis in hypothalamic neuronal cells during oxygen-glucose deprivation[J]. Endocrinology, 2007, 148(1):148-159.
  16. 16. Miao Y, Xia Q, Hou Z, et al. Ghrelin protects cortical neuron against focal ischemia/reperfusion in rats[J]. Biochem Biophys Res Commun, 2007, 359(3):795-800.
  17. 17. Thorens B, Wu YJ, Leahy JL, et al. The loss of GLUT2 expression by glucose-unresponsive beta cells of db/db mice is reversible and is induced by the diabetic environment[J]. J Clin Invest, 1992, 90(1):77-85.
  18. 18. Bell GI, Kayano T, Buse JB, et al. Molecular biology of mammalian glucose transporters[J]. Diabetes Care, 1990, 13(3):198-208.
  19. 19. Fukumoto H, Kayano T, Buse JB, et al. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues[J]. J Biol Chem, 1989, 264(14):7776-7779.
  20. 20. Thorens B, Cheng ZQ, Brown D, et al. Liver glucose transporter:a basolateral protein in hepatocytes and intestine and kidney cells[J]. Am J Physiol Cell Physiol, 1990, 259(6 Pt 1):C279-C285.
  21. 21. Thorens B, Sarkar HK, Kaback HR, et al. Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney, and beta-pancreatic islet cells[J]. Cell, 1988, 55(2):281-290.
  22. 22. Gould GW, Thomas HM, Jess TJ, et al. Expression of human glucose transporters in Xenopus oocytes:kinetic characterization and substrate specificities of the erythrocyte, liver, and brain isoforms[J]. Biochemistry, 1991, 30(21):5139-5145.
  23. 23. Orci L, Thorens B, Ravazzola M, et al. Localization of the pancreatic beta cell glucose transporter to specific plasma membrane domains[J]. Science, 1989, 245(4915):295-297.
  24. 24. Guillam MT, Hümmler E, Schaerer E, et al. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2[J]. Nat Genet, 1997, 17(3):327-330.
  25. 25. Guillam MT, Dupraz P, Thorens B. Glucose uptake,utilization,and signaling in GLUT2-null islets[J]. Diabetes, 2000, 49(9):1485-1491.
  26. 26. Guillemain G, Muñoz-Alonso MJ, Cassany A, et al. Karyopherin alpha2:a control step of glucose-sensitive gene expression in hepatic cells[J]. Biochem J, 2002, 364(Pt 1):201-209.
  27. 27. , 86(10):5083-5086.
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